Hydrolysis of carbohydrate polymers such as cellulose, xylan and starch is used to produce fermentable sugars for bio-ethanol production. Digestion of biomass material and its fermentation represents the best path forward for bio-ethanol production. The use of lignocellulosic material to produce ethanol presents a number of challenges relative to the use of starch containing sources such as corn, which can be digested with amylases. In order to improve the ethanol yield from these polymers, scientists have turned to the use of specific enzymes from a number of different sources. Xylanase enzymes digest xylan polymers, which are a major constituent of the hemicellulose, into xylose.
Optimization of these enzymes in a laboratory or industrial setting requires experimentation with a number of different variables. EnzCheck Ultra Amylase assay (cat # E33651), Amplex® Red Glucose assay (cat# A13261), and EnzCheck Ultra Xylanase assay (cat# E33650) kits as well as the fluorescent cellulase substrate (cat# E33953) were obtained from Life Technologies. Xylanase activity was measured using an EnzCheck Ultra Xylanase assay kit as described previously [2]. Cellulase activity was also measured directly using an EnzCheck Cellulase substrate as described previously [2].
In order to demonstrate the utility of microplates and microplate readers, three different commonly used enzymes responsible for the digestion of polysaccharides used as feed stocks for ethanol production were investigated for activity under a number of environmental conditions.
The effect of pH activity was tested on amylase enzyme isolated from three different sources.
Xylanase enzyme isolated from the fungi Thermomyces lanuginosus was found to be very temperature stable. Note that the effect of pH on the fluorescence of the reacted substrate must be accounted for when comparing results at different pH levels. Enzyme titrations performed at different pH levels corroborate the fixed concentration data.
These data demonstrate the utility of microplate readers to perform classic enzymology experiments on critical enzymes necessary for the digestion of polysaccharide feed stock into fermentable monosaccharides for the production of ethanol.
The use of cellulosic second-generation material to generate fermentable sugars has some marked advantages over first generation starch based grains. The hydrolysis of xylan results in the pentose sugars xylose and arabinose which are not fermented by wild strains of the yeast Saccharomyces cerevisiea.
The use of ethanol as a means to replace a portion of the transportation fuel used in the United States has had legislative support for a number of years. BioTek Instruments, Inc., headquartered in Winooski, VT, USA, is a worldwide leader in the design, manufacture, and sale of microplate instrumentation and software. We also have Bitesize study guides covering many subjects at National 4 and National 5 on our Knowledge & Learning BETA website. In other words, sugar is directly converted to fat by your body, and the more sugar you eat, the fatter you will be.
Consumption of foods that contain large amounts of sugar is a major cause of overweight and obesity. Today I am going to give you a short explanation on how the Glycemic Index works since some of my Clients had brought it out to me when I always talk to them about  Carbs and how, when or how much to add to their Diets. The glycemic index  or GI is a measure of the effects of carbohydrates on blood sugar levels. I personally believe that taking in consideration Glycemic Index goes beyond counting calories since it encourages you to look at the way foods are digested and metabolized in your body and what impact that has on your body weight and how full you feel after eating.
In conclusion (and I hope I am making it easy to understand), Blood sugar, carbohydrates, and insulin all come together to affect body weight, right?
So, even though we may have eaten a high calorie meal, we are induced to feel hungry and eat again within  a very short time. Therefore I insist, instead of counting Calories like crazy, Why not focusing on what type of food you are putting in your mouth and what is the effect that is making in your body?
Recent Content TagsTRX in Smyrna Boot Camp Smyrna Stay fit after 50's GA Low Calories sugar Zumbatomic Atlanta Zumba for kids diet protein lean body How to Get Rid of Flabby Arms? Zumba®, Zumba Fitness® and the Zumba Fitness logos are registered trademarks of Zumba Fitness, LLC. We follow rigorous procedures to qualify manufacturing .this assures we provide our customers quality products that give us the right to give our clients limited warranty.
Get FREE access to authoritative breaking news, videos, podcasts, webinars and white papers. Emsland Group’s Empro® E 86 is a clean label pea protein isolate with an excellent amino acid profile. By admin On October 21, 2011 · 40 Comments In Part 1, we began the process of distinguishing the difference between a food group and a macronutrient.
If we listed the many staple foods: grains, rice, beans, squash, quinoa, potatoes and corn we see a high amount of starch. Sugars are carbohydrates, the biological macromolecules which are most commonly associated with sweetness. In this chapter you will know about the significance of macromolecules such as carbohydrates, proteins, enzymes, lipids and nucleic acids. List the four main classes of large biological molecules as carbohydrates, lipids, proteins and nucleic acids. Classify the carbohydrates based on number of carbons, number of monomers, functional groups, biological function and reducing behavior.


Classify the proteins based on their metabolic function and examine the structure of proteins. Macromolecules are in four main classes: carbohydrates, lipids, proteins and nucleic acids. Each type of small molecule has unique properties arising from the orderly arrangement of its atoms. Small organic molecules are joined inside cells, forming larger molecules called macro molecules. The four main classes of large biological molecules are carbohydrates, lipids, proteins and nucleic acids.
25‐20 percent of living matter consists of macromolecules, including proteins, polysaccharides and DNA.
Though all of these groups are organized around carbon, each group has its own special structure and function.
Just as factories need machines, the machines of living organisms are specific protein molecules called enzymes. The exogenous digestion of these polymers on an industrial scale can be cost prohibitive without optimization of the enzyme reaction conditions.
While starch based feedstocks have been used extensively, it is believed that lignocellulosic energy sources offer a better long-term prospect as a source for fermentable sugars. Using a combination of in vitro digestion, in conjunction with in vivo genetic manipulation of yeast and bacteria strains, the digestion efficiency to fermentable sugars cannot only be increased, but the ability to ferment monosaccharides other than glucose can be improved. The ability to measure multiple samples rapidly in small volumes offers significant advantages in terms of time, expense and effort.
Carboxymethyl cellulose (cat # 419273), xylanase from Thermomyces lanuginosus (cat# X2753), from Trichoderma longibrachiatum (cat # X2629), cellulase from Aspergillus niger (cat# C1184), α-amylase from Aspergillus oryzae (cat# 10065) from Bacillus licheniformis (cat# A4551) and Bacillus subtilis (Cat#10069) were purchased from Sigma-Aldrich. In these experiments, equivalent concentrations of a reference standard were used to normalize data prior to plotting. The direct assay has similar detection limits as the glucose determination, but does not require an overnight digestion. Cellulosic materials do not compete directly with food stocks for resources such as tillable land, fertilizers and water. Its linear chains of glucose moieties closely pack with one another forming hydrogen bonds between chains resulting in a crystalline material that is insoluble in water and many organic solvents. To address this limitation several different strains have been genetically modified in regards to xylanase and arabinose transport proteins and pentose fermentation pathways [4]. These technologies are used to aid life science research, facilitate drug discovery, provide rapid and cost-effective analysis, and enable sensitive, accurate quantification of molecules across diverse applications.
Not surprisingly you will know that almost half of the added sugar in your diet comes from sweetened drinks. As it’s shows in the picture above, The glycemic index of a food is a ranking (from 0 to 100) of how much it raises blood glucose level after it is eaten. High glycemic food cause a larger rise in blood glucose, which can last for a longer time as well.
Carbohydrates that break down quickly during digestion, releasing glucose rapidly into the bloodstream, have a high GI; On the other side, the carbohydrates that break down more slowly, releasing glucose more gradually into the bloodstream, are the ones that have a low GI. So a food with a glycemic index of 95 raises blood sugar almost as much as pure glucose, but a food with a glycemic index of 20 doesn’t raise blood sugar much at all.
This process may lead to excessive calorie intake and weight gain, possibly causing obesity. I once recommended “one serving of protein and one serving of carbohydrate with every meal.” I lost 50 lbs on that advice and yet when challenged by a 11 year old girl with a simple question my “belief system” was stopped in it’s tracks.
What we’ve discussed  is that carbohydrates come in the form of simple sugars (monosaccharides) and more complex carbohydrates (polysaccharides).
We are going to talk about chains, not of steel, but of the basic energy units that keeps your body going and make you fat. We have previously shown the ability to quantitate these enzymes; here we describe the optimization of some of the reaction conditions for the enzymes that catalyze the digestive hydrolysis of polysaccharide polymers into monomeric constituents. Lignocellulosic feedstocks, such as wood chips, corn stover or switchgrasses do not directly compete with food sources for land or consumption. The chain structure forms a stiff extended conformation that results in extensive hydrogen bonding with adjacent chains.
Termites and ruminant herbivores digest cellulose through the action of symbiotic bacteria located in their intestines and ruminating chambers respectively. The use of microplates is one way that large numbers of samples can be handled with minimal amounts of reagents and time.
Amylase enzyme isolated from different organisms were compared for their activity at different pH levels. Enzyme concentration titration curves indicate that higher temperatures result in slightly more hydrolysis than ambient temperatures (Figure 7).
Using the direct substrate assay, the effect of pH on Aspergillus niger cellulase enzyme activity was investigated.
Grain based feed stock rich in starch requires amylase enzymatic activity, while plant and wood material needs cellulase and xylanase activity to break down the cellulose and xylan material contained in the plant fibers. It can be chemically digested with a combination of high temperature and concentrated acids [3]. Ethanol is currently used as an additive to gasoline in the US to replace MBTE as a means to increase gasoline oxygen content.


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So, Rising levels of blood sugar cause the pancreas to produce insulin and Higher levels of insulin then promote body fat storage. He added: “The strategy of our company is to have at least three legs, and one leg is currently a little weaker. This is probably due to the ubiquitous availability of starch foods throughout human history. Regardless of the source of feed stock, current production costs make them unpalatable as a fuel.
Primary and secondary walls contain cellulose, hemicellulose and pectin, albeit in different proportions. The result is the formation of microfibrils with high tensile strength and poor water solubility.
Xylose is a five carbon monosaccharide that can form either furanose (5-member) or pyranose (6-member) ring structures. Here we describe the use of microplates to assess physical assay characteristics, such as pH, temperature and enzyme concentration, of some of the enzymes that catalyze the hydrolysis of plant polysaccharides.
Correction for the influence of pH on fluorescent signal data was made by normalization each well of the initial unreacted read to a common value. Interestingly, temperatures very near ambient (25° C) had the greatest reduction in enzyme activity (Figure 4) when assessed using the fluorescent amylase substrate.
The fluorescence was plotted after 30 minute incubation from assays run at different temperatures. As seen in Figure 10, Aspergillus niger cellulase exhibits significant activity in acidic pH levels. Regardless of the enzymatic requirements it is important that the conditions for maximal enzymatic activity be optimized. However, this technology is currently unfeasible from both an economic and hazardous waste standpoint.
This signals our body to produce insulin, a hormone, so that cells can take the glucose out of the bloodstream and use it for energy. Remember a large quantity of insulin rapidly mops up the excess sugar in our bloodstream causing our blood sugar levels to dip quickly below normal, causing us to feel hungry once more. Polysaccharides are many of these monosaccharides linked together in a chain and are a common way plants store energy (similar to our fat) for later use.
The secondary walls of woody tissue and grasses are composed predominantly of cellulose, lignin, and hemicellulose. In order to correct for the influence of pH on the fluorescent signal data was normalized using a fluorescence reference supplied by the assay kit at each pH.
The effect of temperature was measured by running separate assays at defined temperatures using the Synergy H4 Hybrid Multi- Mode Microplate Reader to maintain temperature. Subsequent data was normalized using the same factor and expressed as the change in signal of the initial unreacted fluorescence. Because microplates provide the ability to test large numbers of samples they are an ideal tool to test multiple experimental conditions.
Towards that end, efforts have focused on ways to reduce the amount of acid used as a treatment by using a combination of dilute acids in conjunction with enzymatic hydrolysis. Because it is currently being used in large quantities the infrastructure to deliver the fuel already exists. The effect of temperature was measured by running separate assays at defined temperature settings using the Synergy™ H4 Hybrid Multi-Mode Microplate Reader to control temperature. Fluorescence for all experiments was measured kinetically with a Synergy H4 Hybrid Multi-Mode Microplate Reader.
Enzymes that can be shown to work well in high temperatures in an acidic environment would provide a distinct advantage over those that are labile under the same conditions. If there is still excess glucose after maxing out glycogen storage, it will be converted and stored as body fat. Cross-linking of this network is believed to result in the elimination of water from the wall and the formation of a hydrophobic composite that limits accessibility of hydrolytic enzymes and is a major contributor to the structural characteristics of secondary walls. As demonstrated in Figure 6, enzymes from these different sources have markedly different responses to differing pH levels.
Likewise the xylanase enzymes that work effectively under similar conditions are desirable as xylan is a significant component of most cellulosic feed stocks.
While you will be able to view the content of this page in your current browser, you will not be able to get the full visual experience. Xylan, which accounts for up to 30% of the mass of the secondary walls in wood and grasses, contributes to the recalcitrance of these walls to enzymatic degradation. Please consider upgrading your browser software or enabling style sheets (CSS) if you are able to do so.
The background fluorescence from an unreacted sample (no enzyme) at that pH raw was first subtracted and then the blanked data normalized by a known fluorescence reference at the same pH. Adenosine diphosphate glucose-starch glucosyltransferase, adenosine diphosphate glucose-starch glucosyltransferase.




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